In this paper, we describe further characterization of the membrane-associated molecule CD26/dipeptidyl peptidase IV (DPP IV), which is said to be adenosine deaminase-binding protein (ADA-bp) in humans, to clarify its association with ADA in rat immune cells. For this purpose, we used three types of rats: DPP IV+ rats; DPP IV- rats, which lack enzyme activity and immunological reactivity of DPP IV; and ADA- rats, which have reduced ADA activity due to continuous peritoneal injection of 2'-DCF, a potent inhibitor of ADA. ADA existed in the cells of DPP IV+ and DPP IV- rats, but it did not exist on the cell surface in either rat. ADA- rats showed a decrease in ADA activity and in the number of immune cells, but no effect on DPP IV was observed. These data suggest that in rats, in contrast to humans, DPP IV does not exist as ADA-bp.